Objective:To clone and express Rv3265c gene of Mycobacterium tuberculosis in Escherichia coli (E.coli) under optimistic conditions,obtain and identify protein expressed,analyze the structure and characteristics of the...Objective:To clone and express Rv3265c gene of Mycobacterium tuberculosis in Escherichia coli (E.coli) under optimistic conditions,obtain and identify protein expressed,analyze the structure and characteristics of the protein using bioinformatics methods for future applications.Methods: Rv3265c gene from Mycobacterium tuberculosis H37Rv was amplified by polymerase chain reaction,and was cloned into the pET-30a vector after purification and recovery.The recombinant plasmid was sequenced and expressed in E.coli BL21(DE3),and then purified and identified by western blotting.The essential physical-chemical properties of the protein were predicated by bioinformatics tools,including subcellular location,secondary structure,domains,antigenic epitopes,etc.Tertiary structure of the protein based on homology modeling was estabUshed,while multi-sequence homological alignment and phylogenetic analysis were preformed.Results:The recombinant protein was obtained in soluble fraction from expression system in E.coli B121(DE3) carrying pET30- Rv3265c plasmid,and Rv3265c gene was expressed correctly.Bioinformatics analysis showed the protein contained no signal peptide and transmembrane helices,located outside of membrane.Secondary structure analysis revealed it containedα-helix,extended strand and random coil,46.8%,14.6%,38.6%,respectively.Furthermore,it possessed six potential antigenic epitopes,one glycosyl transferase domain.A simple three-dimensional model of this protein was constructed by Swiss-model sever.Both sequences and structures were conservative and especial either in gene or in protein.Conclusions:Rv3265c gene might be a desirable molecular target for anti-tuberculosis drug and vaccine.The purified protein from expression will be utilized to study the kinetics of L-rhamnosyltransferase and to develope an enzyme assay for screening vaccine or drug.展开更多
Objective:To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis（MTU）,express the encoded fusing protein in Escherichia coli（E.coli）,identify protein acquired,and predict the structure and function of the p...Objective:To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis（MTU）,express the encoded fusing protein in Escherichia coli（E.coli）,identify protein acquired,and predict the structure and function of the protein utilizing methods of bioinformatics.Methods:fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR.The fbpB-esxA fusing gene Iigated by（Gly<sub>4</sub>Ser）<sub>3</sub> linker was gained by means of Gene Splicing by Overlapping Extension PCU（SOEPCR）, and fusing gene was cloned into expression vector pET-30a.The recombinant plasmid was sequenced and expressed in E.coli BL21（DE3）.The protein was identified by Western blot using anti-HIS antibody.Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics.Results:The UNA sequences fbpB-esxA were identical with that published by GenBank.The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E.coli B1.21（DE3） under the induction of IPTG.Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes.Conclusions:The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag.Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly.The existence of linker doesn’t affect immunogenicity of Ag85B and ESAT-6.It will allow lor characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.展开更多
Three-dimensional( 3 D) fabric composite is a newly developed sandwich structure,consisting of two identical parallel fabric decks woven integrally and mechanically together by means of vertical woven fabrics. In this...Three-dimensional( 3 D) fabric composite is a newly developed sandwich structure,consisting of two identical parallel fabric decks woven integrally and mechanically together by means of vertical woven fabrics. In this paper,six types of 3 D fabric sandwich composites were developed in terms of compressive and flexural properties as a function of pile height( 10, 20 and30 mm) and pile distance( 16, 24 and 32 mm) in pile structures. The mechanical characteristics and the damage modes of the 3 D fabric sandwich composites under compressive and flexural load conditions were investigated. Besides,the influence of pile height and pile distance on the 3 D fabric sandwich composites mechanical properties was analyzed. The results showed that the compressive properties decreased with the increase of the pile height and the pile distance. Flexural properties increased with the increase of pile height, while decreased with the increase of pile distance.展开更多
Favorable physicochemical properties and unique molecular recognition capability endow triptycenebased materials with good potential as stationary phases for gas chromatography (GC). This work reports a new type of tr...Favorable physicochemical properties and unique molecular recognition capability endow triptycenebased materials with good potential as stationary phases for gas chromatography (GC). This work reports a new type of triptycene-based materials functionalized by three benzimidazolium cations with different peripheral alkyl lengths (denoted as TP-3Bim-5C and TP-3Bim-12C) and their GC separation performance. As a result, they shared high resolving performance for the naphthalene isomers but differed for the benzene derivatives with varying polarity. Moreover, their capillary columns exhibited good repeatability and thermal stability. This work presents a facile strategy for tailoring the selectivity of the TP-based stationary phases and demonstrates their promising future for chromatographic analysis.展开更多
A series of soy protein isolate(SPI)films plasticized by glycerol(Gly)were studied using attenuated total reflectance-Fourier transform infrared spectroscopy(ATR/FTIR).Perturbation-correlation movingwindow two-dimensi...A series of soy protein isolate(SPI)films plasticized by glycerol(Gly)were studied using attenuated total reflectance-Fourier transform infrared spectroscopy(ATR/FTIR).Perturbation-correlation movingwindow two-dimensional(PCMW2D)and two-dimensional correlation(2DCOS)analyses were applied to the amideⅠband and thus the hydrogen bond interaction between SPI and Gly was systematically investigated.When Gly concentrations were in the range 0~35%,the hydrogen bond amongβ-sheets was replaced by the one between SPI chain and Gly molecule,which caused these protein chains being changed toα-helix.However,the transformation ofβ-sheet toα-helix was saturated and both of them tend to change to random coil when Gly concentrations were in the range 35%~60%.展开更多
Titania microspheres were synthesized using hydrothermal methods to exploit a new liquid chromatography stationary phase. The prepared titania microspheres were approximately 7 μm in diameter, and the particle size d...Titania microspheres were synthesized using hydrothermal methods to exploit a new liquid chromatography stationary phase. The prepared titania microspheres were approximately 7 μm in diameter, and the particle size distribution was relatively narrow and uniform. Furthermore, the average specific surface area was 276.0 m2·g·1, the average pore volume was approximately 0.25 mL·g·1, and the pore diameter was approximately 35.9 nm for sintering titania microspheres. These parameters indicate that the titania microspheres prepared for this study have excellent surface properties for chromatography. Additionally, columns filled with the titania microspheres were able to separate basic compounds, including benzene, nitrobenzene and o-nitroanisole. It could be proposed that the titania microspheres prepared for this study would be a promising stationary phase for liquid chromatography.展开更多
The unified form of retention equation of chromatography is suggested by adoptingstatistical thermodynamics and lattice theory. According to different conditions, the retentionformulas of GC, LC and SFC can be obtaine...The unified form of retention equation of chromatography is suggested by adoptingstatistical thermodynamics and lattice theory. According to different conditions, the retentionformulas of GC, LC and SFC can be obtained from this unified equation. Those formulashave been confirmed by experimental data.展开更多
基金Supported by Research Program in higher educational institutes of The Education Department of Hainan Province(No.Hj2010-21)Research Program of The Health Department of Hainan Province(No.2007-44)+1 种基金Natural Science Fund of Hainan Province(No.2008-30837)Cultivation Program of Hainan Medical University (HY2010-006)
文摘Objective:To clone and express Rv3265c gene of Mycobacterium tuberculosis in Escherichia coli (E.coli) under optimistic conditions,obtain and identify protein expressed,analyze the structure and characteristics of the protein using bioinformatics methods for future applications.Methods: Rv3265c gene from Mycobacterium tuberculosis H37Rv was amplified by polymerase chain reaction,and was cloned into the pET-30a vector after purification and recovery.The recombinant plasmid was sequenced and expressed in E.coli BL21(DE3),and then purified and identified by western blotting.The essential physical-chemical properties of the protein were predicated by bioinformatics tools,including subcellular location,secondary structure,domains,antigenic epitopes,etc.Tertiary structure of the protein based on homology modeling was estabUshed,while multi-sequence homological alignment and phylogenetic analysis were preformed.Results:The recombinant protein was obtained in soluble fraction from expression system in E.coli B121(DE3) carrying pET30- Rv3265c plasmid,and Rv3265c gene was expressed correctly.Bioinformatics analysis showed the protein contained no signal peptide and transmembrane helices,located outside of membrane.Secondary structure analysis revealed it containedα-helix,extended strand and random coil,46.8%,14.6%,38.6%,respectively.Furthermore,it possessed six potential antigenic epitopes,one glycosyl transferase domain.A simple three-dimensional model of this protein was constructed by Swiss-model sever.Both sequences and structures were conservative and especial either in gene or in protein.Conclusions:Rv3265c gene might be a desirable molecular target for anti-tuberculosis drug and vaccine.The purified protein from expression will be utilized to study the kinetics of L-rhamnosyltransferase and to develope an enzyme assay for screening vaccine or drug.
基金Supported by Research Program of The Health Department of Hainan Province(No.2007-44)Research Cultivation Program of Hainan Medical University(HY2010-006)+1 种基金Research Program in higher educational institutes of The Education Department of Hainan Province(No.Hj2010-21)Natural Science Fund of Hainan Province(No.2008~30837)
文摘Objective:To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis（MTU）,express the encoded fusing protein in Escherichia coli（E.coli）,identify protein acquired,and predict the structure and function of the protein utilizing methods of bioinformatics.Methods:fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR.The fbpB-esxA fusing gene Iigated by（Gly<sub>4</sub>Ser）<sub>3</sub> linker was gained by means of Gene Splicing by Overlapping Extension PCU（SOEPCR）, and fusing gene was cloned into expression vector pET-30a.The recombinant plasmid was sequenced and expressed in E.coli BL21（DE3）.The protein was identified by Western blot using anti-HIS antibody.Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics.Results:The UNA sequences fbpB-esxA were identical with that published by GenBank.The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E.coli B1.21（DE3） under the induction of IPTG.Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes.Conclusions:The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag.Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly.The existence of linker doesn’t affect immunogenicity of Ag85B and ESAT-6.It will allow lor characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.
基金National Key R&D Program of China(Nos.2016YFB0303104,2018YFC0810306)Jiangsu Overseas Visiting Scholar Program for University Prominent Younge Middle-aged Teachers and PresidentsSix Talent Peaks Project in Jiangsu Province,China(No.XCL-061)
文摘Three-dimensional( 3 D) fabric composite is a newly developed sandwich structure,consisting of two identical parallel fabric decks woven integrally and mechanically together by means of vertical woven fabrics. In this paper,six types of 3 D fabric sandwich composites were developed in terms of compressive and flexural properties as a function of pile height( 10, 20 and30 mm) and pile distance( 16, 24 and 32 mm) in pile structures. The mechanical characteristics and the damage modes of the 3 D fabric sandwich composites under compressive and flexural load conditions were investigated. Besides,the influence of pile height and pile distance on the 3 D fabric sandwich composites mechanical properties was analyzed. The results showed that the compressive properties decreased with the increase of the pile height and the pile distance. Flexural properties increased with the increase of pile height, while decreased with the increase of pile distance.
基金the financial support by the National Natural Science Foundation of China(No.21575013)Analysis & Testing Center, Beijing Institute of Technology
文摘Favorable physicochemical properties and unique molecular recognition capability endow triptycenebased materials with good potential as stationary phases for gas chromatography (GC). This work reports a new type of triptycene-based materials functionalized by three benzimidazolium cations with different peripheral alkyl lengths (denoted as TP-3Bim-5C and TP-3Bim-12C) and their GC separation performance. As a result, they shared high resolving performance for the naphthalene isomers but differed for the benzene derivatives with varying polarity. Moreover, their capillary columns exhibited good repeatability and thermal stability. This work presents a facile strategy for tailoring the selectivity of the TP-based stationary phases and demonstrates their promising future for chromatographic analysis.
文摘A series of soy protein isolate(SPI)films plasticized by glycerol(Gly)were studied using attenuated total reflectance-Fourier transform infrared spectroscopy(ATR/FTIR).Perturbation-correlation movingwindow two-dimensional(PCMW2D)and two-dimensional correlation(2DCOS)analyses were applied to the amideⅠband and thus the hydrogen bond interaction between SPI and Gly was systematically investigated.When Gly concentrations were in the range 0~35%,the hydrogen bond amongβ-sheets was replaced by the one between SPI chain and Gly molecule,which caused these protein chains being changed toα-helix.However,the transformation ofβ-sheet toα-helix was saturated and both of them tend to change to random coil when Gly concentrations were in the range 35%~60%.
文摘Titania microspheres were synthesized using hydrothermal methods to exploit a new liquid chromatography stationary phase. The prepared titania microspheres were approximately 7 μm in diameter, and the particle size distribution was relatively narrow and uniform. Furthermore, the average specific surface area was 276.0 m2·g·1, the average pore volume was approximately 0.25 mL·g·1, and the pore diameter was approximately 35.9 nm for sintering titania microspheres. These parameters indicate that the titania microspheres prepared for this study have excellent surface properties for chromatography. Additionally, columns filled with the titania microspheres were able to separate basic compounds, including benzene, nitrobenzene and o-nitroanisole. It could be proposed that the titania microspheres prepared for this study would be a promising stationary phase for liquid chromatography.
文摘The unified form of retention equation of chromatography is suggested by adoptingstatistical thermodynamics and lattice theory. According to different conditions, the retentionformulas of GC, LC and SFC can be obtained from this unified equation. Those formulashave been confirmed by experimental data.
